Journal: BMC Immunology

Article Title: NLRP3 inflammasome up-regulates major histocompatibility complex class I expression and promotes inflammatory infiltration in polymyositis

doi: 10.1186/s12865-022-00515-2

Figure Lengend Snippet: LPS/ATP stimulation activates NLRP3/caspase-1/IL-1β axis and promotes MHC-I expression. A Western blot analysis showed increased expression of NLRP3, pro-caspase-1, cleaved caspase-1, pro-IL-1β and mature IL-1β in the Raw 264.7 macrophages under LPS/ATP stimulation. B , C ELISA analysis showed that the expression of IL-1β and TNF-α in the Raw 264.7 macrophages were up-regulated by LPS/ATP. D – G ELISA analysis showed elevated levels of IFNs and MCP-1 produced by the stimulated macrophages. H The mRNA expression of MHC-I in the co-cultured C2C12 cells was increased under LPS/ATP stimulation and peaked at 48 h. I , J The protein expression of MHC-I in the co-cultured C2C12 cells was increased under LPS/ATP stimulation and peaked at 72 h. * p < 0.01, ** p < 0.001, compared with control group or compared between the groups connecting with the line

Article Snippet: Or in the co-culture system, the C2C12 cells were treated with additional neutralizing monoclonal antibody of IL-1β (1 μg/ml, clone 7E3, InvivoGen).

Techniques: Expressing, Western Blot, Analysis, Enzyme-linked Immunosorbent Assay, Produced, Cell Culture, Control

Journal: BMC Immunology

Article Title: NLRP3 inflammasome up-regulates major histocompatibility complex class I expression and promotes inflammatory infiltration in polymyositis

doi: 10.1186/s12865-022-00515-2

Figure Lengend Snippet: Genetic knockdown of NLRP3 suppresses MHC-I overexpression in C2C12 cells. A Raw 264.7 macrophages were transfected with siRNA targeting NLRP3, and the transfection efficiency was measured by western blot analysis. B , C Raw 264.7 macrophages pretreated with NLRP3-siRNA were stimulated with 200 ng/ml LPS + 500 μM ATP, and then the expression of MHC-I in the co-cultured C2C12 cells was significantly reduced compared with those pretreated with NC-siRNA, as revealed by western blot. D , E The stimulated Raw 264.7 macrophages were pretreated with NLRP3-siRNA or NC-siRNA, and the co-cultured C2C12 cells were added with neutralizing IL-1β monoclonal antibody. No differences in the expression of MHC-I in the co-cultured C2C12 cells were found between those groups stimulated with 200 ng/ml LPS + 500 μM ATP or with PBS, as showed by western blot. ** p < 0.001, compared with control group; ¶¶ p < 0.001, compared with before treatment

Article Snippet: Or in the co-culture system, the C2C12 cells were treated with additional neutralizing monoclonal antibody of IL-1β (1 μg/ml, clone 7E3, InvivoGen).

Techniques: Knockdown, Over Expression, Transfection, Western Blot, Analysis, Expressing, Cell Culture, Control

Journal: BMC Immunology

Article Title: NLRP3 inflammasome up-regulates major histocompatibility complex class I expression and promotes inflammatory infiltration in polymyositis

doi: 10.1186/s12865-022-00515-2

Figure Lengend Snippet: Pharmacological inhibition of NLRP3 inflammasome using MCC950 effectively suppresses MHC-I overexpression in C2C12 cells. A , B Cell viability assay revealed no significant cytotoxicity of MCC950 to the Raw 264.7 cells and C2C12 cells, at the dosage from 0.01 to 10 μM. C LPS/ATP stimulated Raw 264.7 macrophages were treated by MCC950 with dosage from 0.01 to 10 μM, and the IL-1β level in the cell supernatant was significantly decreased at the dosage of 10 μM. D , E LPS/ATP stimulated Raw 264.7 macrophages were pretreated by 10 μM MCC950, and the MHC-I expression in the co-cultured C2C12 cells was significantly suppressed as showed by western blot. * p < 0.01, ** p < 0.001, compared with control group; ¶¶ p < 0.001, compared with before treatment

Article Snippet: Or in the co-culture system, the C2C12 cells were treated with additional neutralizing monoclonal antibody of IL-1β (1 μg/ml, clone 7E3, InvivoGen).

Techniques: Inhibition, Over Expression, Cell Viability Assay, Expressing, Cell Culture, Western Blot, Control

Journal: BMC Immunology

Article Title: NLRP3 inflammasome up-regulates major histocompatibility complex class I expression and promotes inflammatory infiltration in polymyositis

doi: 10.1186/s12865-022-00515-2

Figure Lengend Snippet: IL-1β is crucial for MHC-I up-regulation in C2C12 cells. A , B Recombinant IL-1β was added into the C2C12 cells. The expression of MHC-I in C2C12 cells peaked when treated by 1 ng/ml IL-1β for 48 h (n = 5). C Raw 264.7 macrophages were transfected with siRNA targeting IL-1β, and the transfection efficiency was measured by western blot analysis. D , E LPS/ATP stimulated Raw 264.7 macrophages were pretreated with IL-1β-siRNA or NC-siRNA, and the MHC-I expression in the co-cultured C2C12 cells was significantly lower with the former than with the latter pretreatment, as analyzed by western blot. * p < 0.01, ** p < 0.001, compared with control group; ¶¶ p < 0.001, compared with before treatment

Article Snippet: Or in the co-culture system, the C2C12 cells were treated with additional neutralizing monoclonal antibody of IL-1β (1 μg/ml, clone 7E3, InvivoGen).

Techniques: Recombinant, Expressing, Transfection, Western Blot, Analysis, Cell Culture, Control

Journal: BMC Immunology

Article Title: NLRP3 inflammasome up-regulates major histocompatibility complex class I expression and promotes inflammatory infiltration in polymyositis

doi: 10.1186/s12865-022-00515-2

Figure Lengend Snippet: MCC950 alleviates the intensity of inflammation and MHC-I expression in PM model rats. A , B The muscle section samples of PM modal rats showed a substantial reduction in muscle histological inflammation scores after treated by MCC950. C – E Serum CRP, CK, and LDH levels were significantly reduced after MCC950 treatment in PM modal rats. F – I The expression of NLRP3, MHC-I, and IL-1β were effectively reduced in the muscle tissue of PM model rats by MCC950 treatment. ** p < 0.001, compared with control group; ¶ p < 0.01, ¶¶ p < 0.001, compared with before treatment. Scale bars, 50 μm (original magnification, ×400)

Article Snippet: Or in the co-culture system, the C2C12 cells were treated with additional neutralizing monoclonal antibody of IL-1β (1 μg/ml, clone 7E3, InvivoGen).

Techniques: Expressing, Tissue, Control

Journal: BMC Immunology

Article Title: NLRP3 inflammasome up-regulates major histocompatibility complex class I expression and promotes inflammatory infiltration in polymyositis

doi: 10.1186/s12865-022-00515-2

Figure Lengend Snippet: The level of inflammation is increased in the PM patients and the NLRP3/caspase-1/ IL-1β axis is active in their muscle samples. A Immunohistochemistry analysis showed higher percentage of CD68+ cells (solid black arrows) in the muscle biopsies in the PM patients than in the controls. B Immunofluorescence staining showed positive MHC-I staining (grade+ ~ ++) in the PM patients, and negative in the controls (grade−). C Immunohistochemistry analysis of the muscle biopsies showed higher expression of NLRP3, caspase-1 and IL-1β (solid black arrows) in the PM patients than in the controls. ** p < 0.001; scale bars, 50 μm (original magnification, ×400)

Article Snippet: Or in the co-culture system, the C2C12 cells were treated with additional neutralizing monoclonal antibody of IL-1β (1 μg/ml, clone 7E3, InvivoGen).

Techniques: Immunohistochemistry, Analysis, Immunofluorescence, Staining, Expressing

Journal: bioRxiv

Article Title: Dysplastic Epithelial Repair Propagates Chronic Pathology Through the Paracrine Transformation of Pulmonary Fibroblasts

doi: 10.64898/2026.04.02.716135

Figure Lengend Snippet: ( A ) Quantified levels of IL-1α from naïve fibroblast (Naïve fibro), inflammatory fibroblast (Inflam fibro), and ectopic basal cell (Basal) conditioned media as determined by ELISA. ( B ) Saa3 and Ccl2 gene expression by RT-qPCR for fibroblasts cultured on 0.5kPa plates and treated with a 5-fold titration of IL-1α added to control media, including the level determined in (A). ( C ) Graphic illustrating the strategy used to deplete conditioned media of endogenous IL-1α with antibodies, magnetic beads, and column-based separation. ( D ) Saa3 and Ccl2 gene expression by RT-qPCR for fibroblasts cultured on 0.5kPa plates in control or conditioned media depleted using IgG (CTRL IgG or CM IgG) or IL-1α nAb (CTRL nAB, CM nAb) and supplemented with additional IL-1α nAb after depletion (+Spike). [(B) and (D)] Data shows mean fold change ± SD and are pooled from 3 independent experiments (n=3 mice). P values were calculated using either (B) a ratio paired t -test or (D) a one-way ANOVA with post-hoc Turkey multiple comparison test: * P < 0.05, ** P < 0.01, **** P < 0.0001, ns: not significant.

Article Snippet: The working concentration for the following reagents can be found in the text and figure legends: Human IL-1RA Recombinant Protein (IL1Ra) (PeproTech®, 200-01RA), Mouse IL-1 alpha Recombinant Protein, (PeproTech®, 211-11A), Mouse IL-1α Neutralizing Antibody (Invivogen, Anti-mIL-1α-mIgG1 clone 6H7, mIL-1α-mab9-02), Recombinant Mouse IgG1 Isotype Control Antibody (Invivogen, Anti-β-Gal-mIgG1 clone T9C6, bgal-mab9-02), Human TGF-β 1 Recombinant Protein (PeproTech®, 100-21), Human GDF15 Recombinant Protein (PeproTech®, 120-28C), Human VEGF Recombinant Protein (PeproTech®, 450-32), Human OPN Recombinant Protein (PeproTech®, 120-35), Murine CXCL1 Recombinant Protein (PeproTech®, 250-11), Human IGFBP3 Recombinant Protein (PeproTech®, 100-08), Human TNFRSF11B Recombinant Protein (PeproTech®, 450-14), Murine CXCL5 Recombinant Protein (PeproTech®, 250-17), and Muring CCL20 Recombinant Protein (PeproTech®, 250-27).

Techniques: Enzyme-linked Immunosorbent Assay, Gene Expression, Quantitative RT-PCR, Cell Culture, Titration, Control, Magnetic Beads, Comparison